6 protocols
Title: Affymetrix CEL analysis. Description:
hybridization protocol
Affymetrix Generic Hybridization
nucleic acid labeling protocol
Labeling of samples was carried out according to the manufacturer’s standard protocol
1. Prepare 0.5x MS media pH5.7 with 0.7% phytoagar

2. Sow approx. 300 sterile seeds on plate

3. Stratify for 4 days at 4oC

4. Place in a growth cabinet set at 21oC short days (8h light, 16 h dark)

5. Harvest plants after 10 days of growth

(Parameters: start time = 10, time unit = days, min temperature = 21, temperature unit = C, media = 0.5xMS)
1. Plant culture plates opened in a fume hood

2. Samples sprayed with either mock or DEX solutions:

Mock=0.05% Silwet L77/0.1% Ethanol

DEX =0.05% Silwet L77/0.1% Ethanol/10uM DEX

3. Allow plants to dry

4. Replace lid and return to growth chamber for 4h or 8h

1. 300-600 mg Arabidopsis shoot tissue ground under liquid nitrogen 2. RNA extracted using QIAGEN - RNeasy Plant Mini Kit accoring to manufacturers instructions 3. RNA eluted in 50ul EB (Parameters: Extracted product = total_RNA, Amplification = PCR)